[四种登革病毒血清型包膜蛋白结构域III（ED III）中和抗体的制备与鉴定]

[Preparation and identification of neutralizing antibodies to envelope protein domain III(ED III) of the four dengue virus serotypes].

作者信息

Ding Xixia, Qiu Liwen, Chen Yue, Wen Kun

机构信息

Division of Laboratory Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.

Division of Laboratory Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China. *Corresponding author, E-mail:

出版信息

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2022 Aug;38(8):748-753.

PMID:35851089

Abstract

Objective To prepare neutralizing monoclonal antibodies (mAbs) against envelope protein domain III of the four dengue virus serotypes (DENV-EDIII) and identify its specificity. Methods BALB/c mice were immunized with recombinant EDIII protein (rDENV-EDIII) of the four DENV serotypes. Hybridoma cells secreting DENV-EDIII antibodies were screened by indirect ELISA. The specificity of positive hybridoma cells were further tested by indirect immunofluorescence assay (IFA). The neutralizing activities of DENV-EDIII mAbs in vitro were determined by the enzyme-linked immunospot microneutralization test (ELISPOT-MNT). Results 6 mAbs specific for the EDIII of the four DENV serotypes and 11 mAbs specific for only one serotype of DENV-EDIII protein were obtained, of which one monoclonal antibody had a balanced strong neutralizing activity against the four DENV serotypes in vitro, and its 50% inhibitory concentrations (IC) for the four DENV serotypes was 0.05 μg/mL, 1.89 μg/mL, 0.02 μg/mL, 3.91 μg/mL, respectively. Conclusion A specific monoclonal antibody against DENV-EDIII is successfully screened and obtained to neutralize the four DENV serotypes.

摘要

目的制备针对四种登革病毒血清型包膜蛋白结构域III（DENV-EDIII）的中和单克隆抗体（mAb）并鉴定其特异性。方法用四种登革病毒血清型的重组EDIII蛋白（rDENV-EDIII）免疫BALB/c小鼠。通过间接ELISA筛选分泌DENV-EDIII抗体的杂交瘤细胞。通过间接免疫荧光试验（IFA）进一步检测阳性杂交瘤细胞的特异性。采用酶联免疫斑点微中和试验（ELISPOT-MNT）测定DENV-EDIII mAb的体外中和活性。结果获得了6种对四种登革病毒血清型的EDIII具有特异性的mAb和11种仅对一种血清型的DENV-EDIII蛋白具有特异性的mAb，其中一种单克隆抗体在体外对四种登革病毒血清型具有平衡的强中和活性，其对四种登革病毒血清型的50%抑制浓度（IC）分别为0.05 μg/mL、1.89 μg/mL、0.02 μg/mL、3.91 μg/mL。结论成功筛选并获得了一种针对DENV-EDIII的特异性单克隆抗体，可中和四种登革病毒血清型。